【摘要】 本研究通过对1982年黑龙江某鹅场发生的鹅细小病毒病疑似病例的肝病变组织病毒分离,进行回顾性研究。将肝脏加PBS研磨后,-70℃/37℃反复冻融三次,除菌过滤后接种12日龄鹅胚。发现病毒对鹅胚的致死时间为5～6d,胚体体表有轻微出血点。鹅胚尿囊液经磷钨酸染色,可见具有典型特征的细小病毒粒子。将该病毒尿囊液在12日龄鹅胚中连续传16代,随着传代次数的增加,胚体多集中在3～4d死亡。病毒传至第三代时死亡胚体头、颈、背部出现较为严重的出血点,体表有水肿。第五代病毒尿囊液(命名为HG5/82)对12日龄鹅胚的ELD50为10-5.92/0.1mL。将20只12日龄雏鹅分为三个试验组和一个阴性对照组,各组分别接种鹅胚尿囊液104.92ELD50、103.92ELD50、102.92ELD50及生理盐水,发现雏鹅接种HG5/82后4d出现轻微腹泻,随后恢复正常。用套式PCR检测雏鹅肛拭子病毒的体外排放情况。发现三个试验组在接种病毒后1～20d用套式PCR均可检测到病毒。对照组及试验组接种后20～56d没有检测到病毒。将套式PCR产物进行序列测定并于参考毒株进行比较,表明该序列具有GPV相应区域的共同分子特征,与GPV参考毒株B的相应序列同源性达93.6%。本研究结果表明,该分离毒株为典型的鹅细小病毒(GooseParvovirus,GPV),对雏鹅的致病力较弱。更多还原

【Abstract】 A field strain of goose parvovirus （GPV） was isolated from gosling hepar of affected young layer.Biological characteristics of the virus was studied by virus pathogenicity to goose embryos,morphological observation by electron microscope, ELD₅₀ to 12_day_old goose embryos and virus inoculate to 12 day_old gosling.It was demonstrated that allantoic fluid of the first passage virus produced goose embryos to death about 5～6 days after inoculation and the pathological change was mild.The lesions of goose embryos were obvious after the third passage and the time to death shorten to almost 3 days after inoculation.The virus showed typical shape shared by members of Parvovirus under electron microscope.The ELD50 of the fifth passage virus （HG5/82） to 12_day_old goose embryos was 10^-5.92/0.1 mL. No typical clinical signs were observed after inoculation of GPV_HG5/82 strain. We detected the GPV in the anal swab of the inoculated gosling with nested_PCR and sequencing the product of PCR.It is showed that the HG5/82 virus can be detected within 20 days after inoculation. Furthermore,the nucleotide identity of the PCR product between HG5/82 and GPV strain B was 93.6 %.更多还原